Project Summary/Abstract Core H - The purpose of the Protein Expression and Proteomics Analysis (PEP) Core is to provide state-of- the-art technology, equipment and expertise for the cloning, expression and purification of recombinant proteins required by the San Diego (SD) CFAR community, as well as the tools and expertise required for molecular analysis of proteins and protein mixtures. In particular, the PEP Core will provide protein expression and analytical tools not available to most laboratories and at a reduced cost affordable for CFAR members. The PEP Core is divided into two units based on services provided: The Protein Expression Unit (Co-Director, John Elder) will aid CFAR members as needed for cloning, expression and/or purification of proteins, based on the needs of the investigator. This unit will also maintain stocks of frequently requested viral and immunological proteins such as HIV gp120 and Gag antigens and will aid in the purification of antibodies, as required. The Proteomics Unit (Co-Director, Bruce Torbett) maintains state-of-the-art equipment and capabilities for protein analysis including nanospray mass spectrometer, HPLC, Typhoon 3-color laser for analysis of protein mixtures, light cycler instrumentation for analyzing protein mixtures, and equipment for 1- and 2-dimensional protein separations. These technologies and provided expertise will aid investigators in quantitative and qualitative analysis of their proteins and protein mixtures from cell samples. In addition, the PEP Core will provide training to CFAR members in all facets of protein expression and analysis offered by the Core, as requested by the community. Information will be provided by both one-on-one interactions, as well as through seminars on various techniques, based on the interests of the members. The long-term goal of the Core is to help facilitate efficient and economical analysis of proteins required by the CFAR membership to aid in the more rapid development of an understanding of the HIV life cycle and the development of intervention strategies against the global spread of HIV.